We present a microchip-based, voltage-addressable on/off valve architecture that is fundamentally consistent with the pressures and solvents employed for high-pressure liquid chromatography. Laser photopatterning of polymer monoliths inside glass microchannels is used to fabricate mobile fluid control elements, which are opened and closed by electrokinetic pressures. The glass substrates and crosslinked polymer monoliths operate in water-acetonitrile mixtures and have been shown to hold off pressures as high as 350 bar (5000 p.s.i.). Open/closed flow ratios of 10(4) to 10(6) have been demonstrated over the pressure range 1.5-70 bar (20-1000 p.s.i.), and the pressure-leak relationship shows the potential for valving control of flow through packed or monolithic chromatography columns. We expect that this valve platform will enable multiplexing of multiple chromatographic separations on single microchips.